Partial Characterization of Hexokinase from Euglena gracilis var. bacillaris*

Abstract

SYNOPSIS. A partially purified hexokinase extracted from Euglena had the following properties: 1) pH optimum 8.0–8.5; 2) an absolute requirement for Mg++ and adenosine triphosphate with Mn++, Co++, Zn++, and Ca++ less active or inactive under the test conditions. Inosine triphosphate could not be substituted for adenosine triphosphate; 3) an apparent Km (glucose) of 5 × 10−4 M; 4) the enzyme was not inhibited by the usual sulfhydryl agents, but was inhibited 24% by adenosine diphosphate. The extract phosphorylated fructose and mannose. It was not clear whether a separate enzyme phosphorylated fructose. Relative rates of phosphorylation, as determined by labile phosphate analyses, were: glucose, 1.00; fructose, 1.19; mannose, 0.56. The partially purified extract also contained phosphohexose isomerase activity; 6‐phosphogluconic acid dehydrogenase was absent. Euglena hexokinase is apparently localized in the cytoplasmic or small‐particle fraction of the cell. In most respects the enzyme seems to resemble the well‐characterized yeast hexokinase rather than the muscle enzyme.